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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    The @journal of eukaryotic microbiology 22 (1975), S. 0 
    ISSN: 1550-7408
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Immediate contact with large volumes of cold 50% (v/v) buffered glycerol preserved typical ameboid shape of Chaos chaos and Amoeba proteus with no visible distortions. These technics allowed determination of the contraction sites in these glycerinated models upon application of ATP-Ca-Mg-solutions. The ectoplasmic tube was the main site of contraction. Preliminary EM investigations revealed thick and thin filaments, associated with the ectoplasmic tube near the plasmalemma, which appeared to be the basis for the contractility of the ectoplasmic tube. There was no predominant contraction of the pseudopodial tips or the endoplasm in these models. The changes of volume were as much as 50%, and in some cases were not accompanied by any change in the length of the ameba; however, lengthwise contractions of the ectoplasmic tube in some amebae occurred to as much as 25%. The data substantiate a basic requirement of the ectoplasmic tube contraction theory of ameboid locomotion.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Molecular and cellular biochemistry 135 (1994), S. 71-78 
    ISSN: 1573-4919
    Keywords: calreticulin ; calcium binding proteins ; endoplasmic reticulum ; calcium storage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract In this paper we review some of the rapidly expanding information about calreticulin, a Ca2+-binding/storage protein of the endoplasmic reticulum. The emphasis is placed on the structure and function of calreticulin. We believe that calreticulin is a multifunctional Ca2+-binding protein and that distinct functional properties of the protein may be localized to each of the three structural domains of calreticulin. Most evidence indicates that calreticulin is a resident endoplasmic reticulum protein. However, it can also be found outside of the endoplasmic reticulum compartment, i.e. in the nuclear envelope, in the nucleus, in the cytotoxic granules in T-lymphocytes and in acrosomal vesicles of sperm cells. The evidence reviewed here clearly suggests that calreticulin has other functions in addition to its role as a Ca2+ storage protein in the endoplasmic reticulum.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 260 (1976), S. 525-526 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Glycerination of Amoeba proteus was carried out in 50% glycerol (v/v), EDTA 5 mM, Tris-HCl buffer 0.01 M, KC1 0.05 M, pH 7.0, at 10 C. It is critical to reduce the medium to a small volume containing a few starved amoebae and then rapidly to apply a large volume of this solution. Glycerination was ...
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-4919
    Keywords: calreticulin ; gene expression ; steroid receptor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Calreticulin is a ubiquitously expressed Ca2+ binding protein of the endoplasmic reticulum which inhibits DNA binding and transcriptional activation by steroid hormone receptors. In this study the effects of calreticulin on tyrosine aminotransferase (TAT) gene expression in cultured McA–RH7777 hepatocytes was investigated. McA–RH7777 cells were stably transfected with calreticulin expression vector to generate cells overexpressing the protein. The transcriptional activity of the TAT gene, which is glucocorticoid–sensitive and cAMP–dependent, was investigated in the mock transfected McA–RH7777 and in cells overexpressing calreticulin (designated McA–11 and McA–17). In the presence of dexamethasone or the cAMP analog (CTP–cAMP) expression of the TAT gene was induced in mock transfected McA–RH7777 cells by approximately 4.5 and 5 fold, respectively. In McA–11 and McA–17 cells, overexpressing calreticulin, glucocorticoi ever, the CTP–cAMP–dependent expression of the TAT gene was not affected. The ability of calreticulin to inhibit glucocorticoid–sensitive TAT gene expression but not the cAMP–dependent expression of the gene suggests that the protein affects specifically the action of transcription pathways involving steroid receptors or transcription factors containing KxFF(K/R)R–like motifs. Calreticulin may play an important role in the regulation of glucocorticoid–sensitive pathway of expression of the hepatocytes specific genes during development.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1573-4919
    Keywords: calreticulin ; retinal pigment epithelium ; heat shock ; differentiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Calreticulin is a major Ca2+ binding protein in the endoplasmic reticulum of non-muscle cells. In this report we show that calreticulin protein is strongly induced by heat shock. Activation and attenuation of the heat shock transcriptional response is caused by heat shock factor that binds to 5'-flanking sequences of heat shock responsive genes, the heat shock element. The smallest stretch of DNA that shows detectable binding of heat shock factor in vitro contains a two-sequence unit nGAAnnTTCn which exists in the 5'-flanking region of calreticulin DNA (5'-gGAAccCAGcgTTC-3'). The present data provide direct evidence that calreticulin expression can be modulated by heat shock. Thus, our results strengthen the hypothesis that calreticulin, in addition to its function as a cellular Ca2+ store, is a multifunctional protein which performs at least some of its functions from the lumen of the ER.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 11 (1988), S. 178-181 
    ISSN: 0886-1544
    Keywords: adhesion ; extracellular matrix ; interference reflection microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In most physiological situations, cell contact with a substratum is mediated by proteins of extracellular matrix. Therefore, an increasing number of cellsubstratum adhesion studies employ substrata covered with one or more proteins of extracellular matrix. To visualize the most adhesive cell structures, focal contacts and focal adhesions, the interference reflection microscopy has been widely used. It has been generally accepted that these strongly adhesive structures can be seen as black streaks in interference reflection microscopy. Calculations are presented herein, which although simplified, suggest that when cells are plated on protein-covered substrata, their focal contacts may not always appear black in interference reflection microscopy.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-0878
    Keywords: Calreticulin ; Endoplasmic reticulum ; Sarcoplasmic reticulum ; Calcium-binding proteins ; Rabbit (Lagomorpha)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Calreticulin was identified in a variety of rabbit tissues by Western blot analysis. Indirect immunofluorescence studies on cultured cells or frozen sections from the corresponding tissues revealed that the protein was distributed to the endoplasmic reticulum or sarcoplasmic reticulum. Calreticulin was found to be an abundant calcium-binding protein in non-muscle and smooth muscle cells and a constitutent calcium-binding protein in cardiac and skeletal muscle. From the immunoblot data, calreticulin may exist as an isoform in rabbit neural retina. The present study establishes the ubiquity of calreticulin in intracellular calcium binding.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 239 (1985), S. 451-454 
    ISSN: 1432-0878
    Keywords: Vinculin ; Spectrin ; Actin ; Epithelial sheet ; Immunofluorescence ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the differentiated pigmented epithelial cells of the retina (RPE) of chick embryos cytoskeletal proteins are found in polygonal rings located in the cell cortex. Within the cortical rings of the RPE cells vinculin and spectrin occupy a characteristic position closest to the plasma membrane; actin is found farther away, while tropomyosin and myosin are located farthest from the plasma membrane. The differences in the distribution of these proteins might reflect the functional specialization of different parts of the cortical ring required to develop and transmit tension from individual cells throughout the entire epithelial sheet.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A new pH indicator, seminaphthofluorescein (SNAFL)-calcein acetoxymethyl ester, was used for intracellular pH (pHi) measurement in living MDCK cells with a laser scanning confocal microscope (LSCM) equipped with an Argon/Krypton laser and dual-excitation and dual-emission (FITC/Texas Red) filter set. SNAFL-calcein excitation maxima are ∼492/540 nm (acid/base) and emission maxima are ∼535/625 nm (acid/base) with a pKa value at ∼7.0. The absorption/emission spectra of SNAFL-calcein indicate that the ratio of emission intensities of its basic/acidic forms is pH dependent. With an Argon/Krypton LSCM, we were able to monitor the acidic and basic forms of this dye simultaneously using dualexcitation (488/568 nm) and dual-emission (525-614 nm/∼615 nm) wavelengths (λs). The simultaneous dual-excitation/emission LSCM system allows for efficient recording of pHi dynamics (time resolution ≍ 1 sec) in living cells. We have analyzed emission stability of the dye at different temperatures (22°C and 37°C) and constant pH, and at the same temperature (22D°C) but various pHs (6.6, 7.0, and 7.4). Bleaching rate is slightly higher at 37°C than that at 22°C. The basic form of the dye (λEm ≍ 625 nm) has a slightly higher bleaching rate than the acidic form (λEm ≍ 535 nm) in standard culture medium (pH 7.3) at either 22°C or 37°C. The pHi in MDCK cells calculated from ratio images (535 nm/625 nm) was 7.19 ± 0.03 (mean ± SEM, n = 20). Calibration experiments show that the useful pH range of SNAFL-calcein appears to be between 6.2 and 7.8, as the dye is difficult to calibrate outside this pH range. © 1995 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 155 (1993), S. 14-26 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In the present study, we have examined the regulation of attachment, onset of proliferation and the subsequent growth, in vitro, of chick retinal pigmented epithelial (RPE) cells as a function of the nature of the substratum and of either the activation or inhibition of protein kinase C (PKC). The RPE cells have an adhesive preference for protein carpets which contain laminin. This preference disappears gradually with time in culture. The adhesion of RPE cells to fibronectin is shown to be a receptor-mediated process which involves the RGD recognition signal. This study also demonstrates that a PKC activator, 12-O-tetradecanoyl-phorbol-13-acetate (TPA), affects RPE cell adhesion in a substratum-dependent manner. Exposure of RPE cells to TPA lowers the cell attachment efficacy to ECM protein substrata but does not affect cell attachment to plastic. The onset of cell proliferation is accelerated by TPA on all of the substrata tested. The minimal duration of an effective TPA pulse exerting a long-lasting influence on RPE cell proliferation is between 1.5 and 3.5 hr. Stimulation of cell proliferation by TPA in long-term cultures is independent of the nature of the growth substratum. The acceleration of the onset of cell proliferation by TPA is sensitive to 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7), an inhibitor of conventional PKC, and thus appears to be dependent on the activation of conventional PKC. H7 also affects cell-cell contacts, causing an alteration in the shape (“squaring”) of RPE cells packed into large colonies. Conversely, the effects of TPA on both the attachment and the long-term proliferation of RPE cells are not dependent a conventional PKC isotype, since H7 cannot abolish the influence of TPA on either process. We conclude that the effect of TPA on long-term proliferation of RPE cells is either dependent on a novel PKC isotype or independent of PKC. © 1993 Wiley-Liss, Inc.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
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